FORMULATION AND EVALUATION OF LEVOCETIRIZINE LOADED

Pandey and Tripathi, IJPSR, 2016; Vol. 7(5): 2242-2251. E-ISSN: 0975-8232; P-ISSN: 2320-5148

International Journal of Pharmaceutical Sciences and Research 2242

IJPSR (2016), Vol. 7, Issue 5 (Research Article)

Received on 21 December, 2015; received in revised form, 15 March, 2016; accepted, 24 April, 2016; published 01 May, 2016

FORMULATION AND EVALUATION OF LEVOCETIRIZINE LOADED MUCOADHESIVE

MICROSPHERES FOR NASAL DELIVERY

Jaideo Pandey *1

and Pushpendra Kumar Tripathi 2

Department of Pharmaceutics 1, R.R.S. College of Pharmacy, Amethi, 227405, Uttar Pradesh, India

Department of Pharmaceutics 2, R.I.T.M., Lucknow, Uttar Pradesh, India

ABSTRACT: The purpose of research work was to develop and optimize

mucoadhesive microspheres of levocetirizine for nasal delivery with the aim to

enhance the residence time and improve therapeutic efficacy and at the same time

increase the Local absorption of drug and reducing systemic side effects and also to

develop unique delivery system for patients suffering from allergy and rhinitis.

Chitosan (mucoadhesive) based microspheres of levocetirizine were prepared by

emulsification-crosslinking method. Glutaraldehyde was used as crosslinking agent.

The mean particle size was significantly increased when high concentration of

chitosan was used. Aqueous to oil phase ratio, stirring rate and dioctyl sodium

sulfosuccinate (DOSS) concentration also influenced the particle size distribution of

the microspheres. Microspheres were evaluated with respect to the production yield,

particle size, entrapment efficiency, swelling index, FT-IR, in vitro mucoadhesion,

% cumulative drug release, histological study and stability studies. Formulation Lf3

was found to be optimized. The optimized formulation Lf3 was mucoadhesive in

nature which adhere onto the mucus and increase the residence time within the nasal

cavity.

INTRODUCTION: The nose is considered as an

attractive route for needle-free vaccination and for

systemic drug delivery, especially when rapid

absorption and effect are desired. In addition, nasal

delivery may help address issues related to poor

bioavailability, slow absorption, drug degradation,

and adverse events in the gastrointestinal tract and

avoids the first-pass metabolism in the liver.

However, when considering nasal delivery devices

and mechanisms, it is important to keep in mind

that the prime purpose of the nasal airway is to

protect the delicate lungs from hazardous

exposures, not to serve as a delivery route for drugs

and vaccines.

QUICK RESPONSE CODE

DOI: 10.13040/IJPSR.0975-8232.7(5). 2242-51

Article can be accessed online on: www.ijpsr.com

DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.7 (5). 2242-51

Nasal drug delivery possesses various advantages

as a site for drug delivery, such as it provides much

vascularized epithelium, large surface area for drug

absorption, lower enzymatic activity compared

with the gastrointestinal tract and liver and the

direct drug transport into the systemic circulation,

thereby avoiding hepatic first pass metabolism and

irritation of gastrointestinal membrane 1. Nasal

route is non-invasive therefore, reduced risk of

infection, ease of convenience and self-medication

resulting in improved patient compliance. The

range of compounds investigated for possible nasal

application greatly from very lipophilic drugs to

polar, hydrophilic molecules including peptides and

proteins 2.

In nasal drug delivery, the most important

limitation factor is rapid mucociliary clearance,

which is the cause of a limited contact period

allowed for drug absorption through the nasal

mucosa. Thus, mucoadhesive nano and micro-

Key words:

Levocetirizine,

Mucoadhesive

Microspheres, Chitosan,

Nasal Delivery, Emulsification

Crosslinking Method

Correspondence to Author:

Mr. Jaideo Pandey

Associate Professor

Department of Pharmaceutics,

R.R.S. College of Pharmacy,

Amethi, 227405, (U.P.) INDIA

Email: [email protected]



particles have been formulated to overcome the

rapid mucociliary clearance, thereby increasing

drug absorption through nasal cavity. Chitosan is a

natural polymer that has mucoadhesive properties

because of its positive charges at neutral pH, which

enable an ionic interaction with the negative

charges of sialic acid residues on the mucus 3. This

highly mucoadhesive characteristics of chitosan

provide a longer contact period for drug transport

through nasal mucosa and prevents the clearance of

the formulation via mucociliary clearance

mechanism 4. Therefore, chitosan microspheres

have been extensively evaluated as a drug delivery

system. In this study, we aimed to formulate

levocetirizine-loaded mucoadhesive microspheres

with chitosan and to investigate feasibility of

levocetirizine nasal delivery with chitosan

microspheres.

Levocetirizine is a third generation antihistamine

acts by blocking histamine receptor. Which is used

in the treatment of allergy & rhinitis5? It is

generally given by oral route. However sometimes

its oral route which makes oral treatment

unsatisfactory. Intranasal route may be viable

alternative for self-administration where the

limitations of oral and parenteral route could be

overcome. Conventional dosage forms may be

unsatisfactory due to their poor residence time in

nasal cavity. Mucoadhesive polymer like chitosan

can be employed to increase the residence time of

the formulation to enhance the bioavailability 6.

Chitosan microspheres have received considerable

attention as nasal drug delivery systems. Chitosan,

being biodegradable, biocompatible, and non-toxic

and bioadhesive polymer. Chitosan is a cationic

polysaccharide, derived by the deacetylation of

chitin. Chitosan is positively charged due to its

amino group and able to interact strongly with the

negatively charged mucus layer of the nasal

epithelium 7.

This is to provide a longer contact time for drug

transport across the nasal membrane, before the

formulation is cleared by the mucociliary clearance

mechanism. In addition, chitosan has been shown

to increase the paracellular transport of polar drugs

by transiently opening the tight junctions between

the epithelial cells. In the present study chitosan

microspheres intended for nasal delivery of

levocetirizine were prepared by emulsification

crosslinking technique using glutaraldehyde (GLA)

as the crosslinking agent 8, 9, 10

. Hence, in the

present work, an attempt was made to formulate

and evaluate mucoadhesive microspheres of

levocetirizine that will increase residence time in

the nasal cavity and at the same time increase the

local of absorption of drug and reducing systemic

side effects and also to develop unique controlled

delivery system for patients suffering from allergy

and rhinitis. The microspheres were prepared by

emulsion cross linking method in different ratio by

using mucoadhesive polymer, chitosan.

MATERIALS:

Levocetirizine was received as a kind gift from

Ajenta Pharma Ltd. (Mumbai, India). Chitosan was

provided by Fisher scientific, Mumbai, India. All

other ingredients used were of analytical grade and

were used without further purification.

Spectrophotometric studies were carried out by

using double-beam UV-spectrophotometer,

Shimadzu, Pharma Spec 1700, Kyoto, Japan.

Methods:

Preparation of mucoadhesive microspheres:

Chitosan microspheres were prepared by simple

w/o emulsification-cross linking process using

liquid paraffin (heavy and light 1:1) as external

Phase6. Briefly, chitosan was dissolved in 2%

aqueous acetic acid solution by continuously

stirring until a homogeneous solution was obtained

(Table 1). Specified quantity of drug dispersed

homogeneously by stirring in chitosan solution.

This solution was added slowly to liquid paraffin

(heavy and light 1:1) containing 0.2% (w/v) of

DOSS as stabilizing agent under constant stirring at

1200 rpm-1375 rpm speed for 15 min using a

Eurostar (IKA Labortechnik, Germany) high speed

stirrer. To this w/o emulsion, Glutaraldehyde

(GLA) was added slowly in definite concentration

(2 ml) in different formulation and stirring was

continued for 2 hrs. The hardened microspheres

were separated by vacuum filtration and washed

several time with hexane to remove oil. Finally,

microspheres were washed with distilled water to

remove unreacted GLA. The microspheres were

dried for 24 hrs and then stored in vacuum

desiccators until further use.



TABLE 1: FORMULATION COMPOSITION OF MUCOADHESIVE MICROSPHERES

Formulation & process variables Constant parameters

Formulations Drug: polymer

ratio

% of

stabilizer

used

(DOSS)

Vol. of cross linking

agent (Glutaraldehyde)

Aqueous to

oil phase

ratio

Stirring

rate

Cross

linking

time

Lf1 1:1

0.2

2ml

10:100

1375 rpm

2 hours

Lf2 1:2

Lf3 1:3

Lf4 1:4

Characterization of levocetirizine loaded

microspheres:

Particle size: 11

, 12

The particles size of the microspheres measured by

using optical microscope(OLYMPUS CH 20i)

Equipped with modified software Magnus pro 3.0

and Olympus master through a camera using a

quantity of microspheres suspended in glycerin

and the mean particle size was calculated by

measuring more than 100 microspheres were

measured randomly by optical microscope.

Production yield: 13

The production yield of microspheres of various

formulation were calculated using the weight of

final product after drying with respect to the initial

total weight of the drug and polymer used for

preparation of microspheres.

Determination of entrapment efficiency: 13

Accurately weighed equivalent to 5 mg of

levocetirizine microspheres were crushed and

dissolved in 100 ml methanol with the help of

ultrasonic stirrer and kept overnight The Solution

was filtered through Whatmann filter paper No.41,

suitable dilution (6,8,10 mcg/ml). The samples

were assayed for drug content by UV-

spectrophotometer at 231.1nm. The drug

entrapment efficiency was calculated using

following Equations (1).

Entrapment efficiency (%) =

Equ……………(1)

Where Mactual is the actual levocetirizine content in

weighed quantity of powder of microspheres and

Mtheoretical is the theoretical amount of levocetirizine

in microspheres calculated from the quantity added.

Scanning Electron Microscopy:

The surface morphology of optimized formulation

(Lf3) was examined by scanning electron

microscopy (JSM 6390, India). The images were

recorded at the 100X magnification at the

acceleration voltage of 10 kv14

.

Fourier Transform Infrared Spectroscopy (FT-

IR spectroscopy):

Levocetirizine, Chitosan and optimized formulation

(Lf3) was examined using FTIR Spectrophotometer

(Shimadzu FTIR-8400S Kyoto, Japan). The test

sample diluted with KBr to get a final dilution of

1:10 was mounted into the instrument. The

measurements were made in transmittance mode in

the range of 400-4000 cm-1

against the background

spectra of pure KBr by setting resolution of 4 cm-1

and 50 times accumulation 15

.

Swelling ability of microspheres: The swelling ability of microspheres was

determined by allowing them to swell to their

equilibrium in phosphate buffer of pH 6.416, 17

.

Swelling was determined in triplicate by using the

equation 2.

Equ………………2

Where α is degree of swelling, Wo is initial weight

of microspheres and Ws is the weight of

microspheres after swelling.

Mucoadhesive Testing by in-vitro wash-off test: In Mucoadhesive properties of the microspheres

were evaluated by in vitro adhesion testing method

known as the wash-off method 18

. In this method

freshly excised nasal mucosal membrane (3×2 cm)

of goat was taken and mounted on the paddle of

USP dissolution test apparatus with thread

Microspheres were spread onto each wet rinsed

tissue specimen, and immediately therefore the



support washing onto the arm of a USP dissolution

test apparatus. Operate USP dissolution test

apparatus at 25 rpm of paddle in phosphate buffer

6.4 at 37°C ± 0.5°C. At the end of 30 min, 60 min,

at hourly intervals up to 6 hours.

In-vitro Release Studies:

The drug release study was performed using USP

XXIV basket apparatus at 37°C ± 0.5°C at 50 rpm

using 900 mL of phosphate buffer (pH 6.4) as a

dissolution medium as per USP XXVI dissolution.

Microspheres equivalent to 5 mg of levocetirizine

drug were used for the test. Five milliliters of

sample solution was withdrawn at predetermined

time intervals, filtered through a Whatmann filter

paper, diluted suitably and analyzed

spectrophotometrically19, 20, 21

. An equal amount of

fresh dissolution medium was replaced

immediately after with drawl of the test sample.

Percentage drug dissolved at different time

intervals was calculated at 230.1 nm.

Kinetics of Drug release:

To examine the drug release kinetics and

mechanism, the cumulative release data were fitted

to models representing zero order (Q v/s. t), first

order [Log (Q0‐Q) v/s. t], Higuchi’s square root of

time (Q v/s. t 1/2) and Korsemeyer Peppas double

log plot (log Q v/s. log t) respectively, where Q is

the cumulative percentage of drug released at time t

and (Q0‐Q) is the cumulative percentage of drug

remaining after time t. In short, the results obtained

from in vitro release studies were plotted in four

kinetics models of data treatment as follows:-

Cumulative percentage drug release Vs.

Time (zero order rate kinetics)

Cumulative percentage drug release Vs. √T

(Higuchi’s classical diffusion equation)

Log cumulative percentage drug release Vs.

log time (Korsmeyer Peppas equation)

Log cumulative percentage drug remaining

Vs. time (First order rate kinetics)

Kinetic analysis was performed and the data was

evaluated after fitting to Zero order, First order,

Higuchi, Peppas values observed where Regression

co-efficient (R) and Diffusion exponent (n) value in

case of Peppas model. Criteria for selecting most

appropriate model were based on best reliability of

fit indicated by ‘R’ value nearer to one. When drug

release is concentration dependent, first order

model is an indicator. Zero order model is

independent of concentration of drug. Matrix

model is applicable when matrix polymer is used

and Peppas model is used when release mechanism

is not well known Fickian diffusion exists when

n<0.5, but at n>0.5 non-fickian diffusion

mechanism was observed 22, 23, 24, 25

Histological studies: Histological studies were conducted to determine

the effect of formulation on nasal mucosa. Nasal

mucosa of Goat was obtained from slaughter house

in saline phosphate buffer pH 6.4.The mucosa was

kept in 10% formalin solution for stabilize the

mucosa. Three pieces of nasal mucosa of identical

size were cut and mounted on separate glass slide.

On one slide was trated with0.5ml phosphate buffer

pH 6.4(negative contol) Second slide treated with

0.5 ml isopropyl alcohol(positive control), in third

slide slide formulation Lf3(control) and all the slide

kept for for 6 h. After 6 h slides were subjected to

histopathology study for evaluation of nasal

toxicity 3, 26, 27

. The specimens were visualized

through Microscope at 100 x magnification at Pt.

Deen Dayal Upadhaya Pashu Chikitsa Vigyan

Vishwavidyalya and Gau research center Mathura,

India.

Stability studies:

The optimized formulation Lf3 was tested for

stability studies. The formulations were divided

into 3 sets of sample and stored at 4±1˚C, 25±2˚C

and 60±5% RH, 37±2˚C and 65±5%RH 28, 29

. After

one to six month, the drug release of selected

formulations was determined by the method

discussed previously in vitro drug release studies

and percentage entrapment efficiency was also

carried out for the same formulation.

RESULT AND DISCUSSION:

Preparation of microspheres:

In the present study, Emulsification-cross linking

method described here approved a suitable and

simple technique to prepare chitosan microspheres

loaded with levocetirizine. For preparation of W/O



type of emulsion, polar organic solvent was

employed as ‘aqueous phase’.

Characterization of levocetirizine loaded

mucoadhesive microspheres:

Particle Size: The mean particle sizes of the

formulations were shown in the table 2. The mean

particle size of microspheres ranged from 11-24

µm. The particle size mainly depends on the

stirring rate and slow effect of concentration of

mucoadhesive polymers, it is clear that stirring rate

increases particle size decreases both at higher and

lower concentration of polymers while

concentration of mucoadhesive polymer had

opposite effect on particle size.

Production yield:

The production yields of microspheres prepared by

emulsion cross-linking method were found to be

between 63.96-75.2% in case of Levocetirizine as

shown in table 2. It was found that production yield

of microspheres prepared by 1:3 (drug: polymer)

was greater than Lf1 (1:1), Lf2 (1:2), and Lf4 (1:4).

The probable reason behind this may be the high

viscosity of the chitosan solution wastage of the

drug-polymer solution which ultimate decreased

the production yields of microspheres. Another

reason for that may be agglomeration and sticking

of polymer to blades of stirrer and to the wall of the

beaker during microsphere formation.

Entrapment efficiency:

Entrapment efficiency was high since it always

exceed 75%. It was found that with increasing the

ratio of drug to polymer, the entrapment efficiency

was also increased (Table 2).

TABLE 2: PARTICLE SIZE OF LEVOCETIRIZINE LOADED FORMULATIONS

Formulation

code

Particle size

(μm)

Production

Yield %

Encapsulation

efficiency %

Mucoadhesion % Swelling index

%

Lf1 24±3.000 63.96±0.451 79.36±0.472 64.83±0.289 0.623±0.008

Lf2 20.33±4.163 68.43±0.404 83.2±0.3 70±0.500 0.828±0.037

Lf3 11±1.000 77.8±0.755 87.2±0.7 76±0.500 0.956±0.050

Lf4 16.19±8.308 75.2±0.300 84.76±0.51 78.03±0.451 1.08±0.076

N = mean of 3, SD±= Standard Deviation

Scanning Electron Microscopy:

The optimized formulation Lf3was examined by

SEM. SEM images of Lf3 in presented in Fig. 1.

SEM analysis revealed that optimized formulation

Lf3 microspheres were spherical in shape and

microspheres have smooth surface.

FIG 1: SEM OF FORMULATION (LF3)



Fourier transforms infrared spectroscopy

(FTIR):

FTIR spectroscopy to know any possible

interaction between levocetirizine, chitosan and the

crosslinking agent. Levocetirizine and chitosan

showed characteristic peak at range of 400-4000

cm-1

. The FTIR spectrum of chitosan in Fig. 2

showed peaks corresponding to O‐H stretching at

3428 cm‐ 1 and amine group (NH2) stretching at

2958.1 cm‐1 respectively. The spectrum of drug

loaded microspheres denotes that the drug was

intact in the formulation and the absence of drug-

polymer interaction. Changes in the intensity of the

peaks indicating no interaction between drug and

polymer.

FIG.2: FTIR SPECTRA OF (a) LEVOCETIRIZINE, (b) CHITOSAN AND (c) LEVOCETIRIZINE LOADED MICROSPHERES

Swelling ability of microspheres:

The swelling index of all formulation was shown in

Table 2. From the table, degree of swelling for

chitosan microspheres varied from 0.623±0.008 to

1.08±0.076. It is known that the degree of swelling

increases marginally as the concentration of

mucoadhesive polymer increases. Marginal

decrease in swelling at lower level of

mucoadhesive polymer may be due to the higher

level of film forming polymer (chitosan) in those

formulations which allows lesser penetration of

water inside the polymer matrix. From this, it may

be concluded that when the microspheres are in

contact with mucus layer, they swell rapidly and

take up liquid from the mucus layer, Hence, the

epithelial cells loose water and shrink which opens

the epithelial tight junctions allowing drug to be

absorbed.

In vitro Mucoadhesion:

The mucoadhesion of levocetirizine loaded nasal

microspheres closely varied between 64.83±0.289



to 78.03±0.451 (Table 2) and was dependent on

polymer concentration. Such excellent

mucoadhesion of chitosan microspheres were from

the electrostatic attraction between chitosan and

mucin. Moreover, the linear molecules of chitosan

expressed sufficient chain flexibility for

interpenetration and entanglement. A good

mucoadhesion is the high flexibility of polymer

backbone structure and its polar functional groups.

Such flexibility of the polymer chain is reduced if

the polymer molecules are cross-linked either with

each other or with cross-linking agent. The

decrease in flexibility imposed upon polymer chain

by cross-linking makes it more difficult for cross-

linked polymer to penetrate the mucin network.

Thus cross-linking effectively limits the polymer

chain that can penetrate the mucus layer and could

possibly decrease mucoadhesion strength.

In vitro release studies: The in vitro release data of all the formulations

were tabulated in Table. The cumulative drug

release after 8hrs was found to be 81% , 81.83%,

86.03%, 82.83% respectively for the formulation

Lf1 to Lf4 (Table 3). The release studies of

Levocetirizine loaded chitosan microspheres are

graphically shown in Fig. 2. It was clear that both

the variables (stirring rate & concentration of

polymer) had significant impact on drug release. As

the concentration of mucoadhesive polymer

increased, the drug release also increased

proportionally. Stirring rate had more influence on

drug release than concentration of mucoadhesive

polymer. Drug release increased steeply as the

stirring rate was increased from lower to higher

level.

This presumably is due to the smaller particle size

of microspheres at higher stirring rate which leads

to much larger surface area available for release

and shorter path length for drug to diffuse through

microspheres. The greater drug release from

chitosan microspheres may be due to the higher

swelling degree of chitosan which forms

hydrophilic passage inside the microspheres who

help drug diffuse out. The increase hydrophilic

pores formed by chitosan facilitated the water

penetrating into microspheres, accelerated the

erosion of swelling matrix and resulted in a

combination of the diffusion and erosion

mechanism of drug release from microspheres.

From the percent drug release graph, formulations

Lf3 were showed best result.

TABLE 3: IN VITRO DRUG RELEASE OF LEVOCETIRIZINE LOADED MICROSPHERES

Time (hrs) Formulation Code

Lf1 Lf2 Lf3 Lf4

0 0 0 0 0

1 14.5±0.500 18.5±0.500 23.16±0.289 22.2±0.721

2 23.7±0.608 23.53±0.500 33.33±0.577 32.5±0.500

3 35.4±0.529 38.83±0.764 43±0.500 42.16±0.764

4 41±0.500 43.33±1.607 51.5±0.500 50.83±1.041

5 51.7±0.265 52.5±0.500 62±0.500 61.4±0.529

6 60.83±0.794 62.83±0.764 68.5±0.500 68.5±0.500

7 68.5±0.500 71±0.500 76±0.500 73±1.000

8 81±0.500 81.83±0.764 86.03±0.451 82.83±0.764

N = mean of 3, SD±= Standard Deviation

FIG. 3: IN VITRO RELEASE OF LEVOCETIRIZINE LOADED MICROSPHERES



In vitro Drug release kinetics studies: The in vitro drug release data of all the

formulations were fit into Zero order, First order,

Higuchi Equation and Korsemeyer-Peppas model.

The results were shown in Table 4. The ‘R’ values

for zero order kinetics of Lf1 to Lf4 were 0.979 to

0.998 and ‘R’ values for first order kinetics of Lf1

to Lf4 were 0.929to 0.970 respectively. Among the

zero order and first order equations, the Zero order

Regression co-efficient (R2) value was found to be

more than the First order. So all the formulations

Lf1 to Lf4 followed Zero order drug release values

indicate the drug release follows zero order (Fig.

4). To ascertain the drug release mechanism, the in-

vitro data were also subjected to Higuchi diffusion.

The ‘R’ values of Higuchi diffusion was 0.945 to

0.965 for formulation Lf1 to Lf4 respectively. So it

confirms the drug release by Higuchi diffusion

mechanism. Higuchi equation explains the

diffusion controlled release mechanism. The

diffusion exponent (n) values of

Korsemeyer‐Peppas model was found to be All the

formulations were subjected to Korsmeyer-Peppas

plots, ‘n’ value ranges from 0.700 to 0.810

indicating that the drug release was by non-fickian

diffusion mechanism (Table 4).

TABLE 4: REGRESSION CO-EFFICIENT (R) VALUES IN THE ANALYSIS OF RELEASE DATA OF MICROSPHERES AS PER

VARIOUS KINETICS MODEL AND DIFFUSION EXPONENT (N) VALUE OF PEPPAS EQUATION

Formulation Code Zero

order

First order Higuchi

matrix

Peppas plot Best fit model

r2 value r

2 value r

2 value r

2

value

‘n’ value

Lf1 0.979 0.929 0.945 0.943 0.810 Zero order

Lf2 0.982 0.939 0.960 0.968 0.792 Zero order

Lf3 0.998 0.970 0.965 0.988 0.744 Zero order

Lf4 0.985 0.945 0.941 0.976 0.700 Zero order

FIG. 4: ZERO ORDER RELEASE KINETICS OF LEVOCETIRIZINE MICROSPHERES FORMULATIONS.

Histological studies:

Nasal mucosa of Goat was obtained from slaughter

house in saline phosphate buffer pH 6.4. The

mucosa was kept in 10% formalin solution for

stabilize the mucosa. Three pieces of nasal mucosa

of identical size were cut and mounted on separate

glass slide. On one slide was treated with0.5ml

phosphate buffer pH 6.4 (negative control) Second

slide treated with0 .5 ml isopropyl alcohol (positive

control), in third slide formulation Lf3 (control)

and all the slide kept for 6 h. After 6 h slides were

subjected to histopathology study for evaluation of

nasal toxicity. The specimens were visualized

through Microscope at 100 x magnification. Nasal

toxicity study was performed to evaluate any toxic

effect of drug and excipients were used in

formulation of microspheres on nasal mucosa. In

negative control treated with phosphate buffer pH

6.4 nasal mucosa appeared intact with no signs of

nasal mucosa damage. While positive control with

isopropyl alcohol shows extensive damage of nasal

mucosa. After treating with microspheres

formulations the nasal mucosa shows no sign of

any damage. Hence the developed microspheres

formulation can be considered as safe for nasal

application (Fig.5).



FIG.5: HISTOLOGICAL STUDY (a) NEGATIVE CONTROL, (b) POSITIVE CONTROL AND (c) FORMULATION (LF3)

Stability studies:

Stability studies of the prepared Levocetirizine

microspheres were carried out by storing the best

formulation Lf3 at 4±1˚C, 25±2˚C & 60±5˚C RH

and 37 ± 2˚C & 65 ± 5% RH for six month.

Parameter namely percentage entrapment

efficiency and percentage cumulative drug release

was carried out. The result of entrapment efficiency

and percentage cumulative drug release after six

months of storage were shown in Table 5. These

studies revealed that, there is a

reduction in entrapment efficiency and percentage

cumulative drug release after six months at 4±1˚C,

25±2˚C & 60±5˚C RH and 37±2˚C & 65±5% RH.

It was also revealed that formulations stored at

25±2˚C& 60±5˚% RH showed maximum

entrapment and percentage cumulative drug release

followed by the storage at 4±1˚C and 37±2˚C;

65±5% RH conditions. These results may be

attributed to erosion of polymer matrix to some

extent during storage (Table 5). TABLE 5: STABILITY STUDIES OF THE OPTIMIZED FORMULATIONS (LF3)

Time in

Month

4±1ºC 25±2ºC &

60±5% RH

37±2ºC & 65±5% RH

EE (%) % CDR EE (%) % CDR EE (%) % CDR

1 87.2 86 87.2 86.03 87.2 86.02

2 87.1 86 87.1 86.02 87 86

3 87 85.9 87.1 86 86.8 85.8

4 86.5 85.8 87.1 86 85.5 85.7

5 86.5 85.7 87 85.9 85 85.7

6 86.2 85.6 87.0 85.9 85.0 85.5

CONCLUSION: In the present studies, it can be

concluded that Levocetirizine microspheres based

on chitosan prepared by emulsification cross

linking method may be considered a promising

nasal delivery. Thus, the formulated microsphere

seems to be potential candidate as intranasal

controlled drug delivery system for the treatment of

allergy & rhinitis.

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How to cite this article:

Pandey J and Tripathi PK: Formulation and Evaluation of Levocetirizine Loaded Mucoadhesive Microspheres for Nasal Delivery. Int J

Pharm Sci Res 2016; 7(5): 2242-51.doi: 10.13040/IJPSR.0975-8232.7(5). 2242-51.

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FORMULATION AND EVALUATION OF LEVOCETIRIZINE LOADED